Targeting to compartments of the endomembrane system for the accumulation of HIV-1 p24 in tobacco plants [Elektronische Ressource] / presented by Maria Goretti Virgili López

Goretti

Sujets

• Biologie

Informations

• Publié par: ruprecht-karls-universitat_heidelberg
• Publié le: 01 janvier 2008
• Nombre de lectures: 36
• Langue: English
• Poids de l'ouvrage: 4 Mo

Extrait

Heidelberg Institute for Plant Science






Targeting to compartments of the endomembrane
system for the accumulation of HIV-1 p24 in
tobacco plants










Maria Goretti Virgili López







Ruprecht-Karls-Universität Heidelberg



Dissertation

Submitted to the

Combined Faculties for the Natural Sciences and for Mathematics
of the Ruperto-Carola University of Heidelberg, Germany

for the degree of
Doctor of Natural Sciences










presented by

Master of Science

Maria Goretti Virgili López
born in
Tarragona, Spain












Oral examination: 24.10.2008













Targeting to compartments of the endomembrane
system for the accumulation of HIV-1 p24 in
tobacco plants











Gutachter: Prof. Dr. David G. Robinson
Prof. Dr. Karin Schumacher











Quando você quer alguma coisa, todo o Universo conspira
para que você realize o seu desejo

When you really want something to happen, the whole universe conspires
so that your wish comes true





É justamente a possibilidade de realizar um sonho que torna a vida
interessante

It’s the possibility of having a dream come true that makes life interesting








Paulo Coelho, The Alchemist



































To my beloved family



Acknowledgments

I am very grateful to many people helping me during my studies. Without them, it is
really hard to imagine how I would have been able to finish my thesis.

First, I wish to thank my three supervisors, Prof. Dr. David G. Robinson, Prof. Dr. Jean-
Marc Neuhaus and Dr. Alessandro Vitale, for giving me such a precious opportunity to
join their labs and for valuable advice, guidance and encouragement.

I would like to thank Prof. Dr. Karin Schumacher, Prof. Dr. Benhard Dobberstein and
Prof. Dr. Ralf Bartenschlager to accept to be the committee of my examination.

I am grateful to the Pharma-planta consortium for granting me with the PhD
scholarship.

I also really appreciate all the useful advice and help from Dr. Guillaume Gouzerh, Dr.
Markus Langhans, Dr. Julia Bubeck, Dr. Maddalena de Virgilio and Dr. Alessandra
Barbante, their encouragement and trust make me feel so lucky to have such wonderful
colleagues.
I also give my thanks to the different lab people I have met in the three laboratories I
have worked. In Neuchâtel: Dr. Ana Slaughter, Dr. Marc Creus, Dr. Anita Ivanova, Dr.
Zhanna Kravchuk, Dr. Ilian Simidjiev, Sophie Marc-Martin, Dr. Laurent Zimmerli, Dr.
Okmeni N’Guemelieu, Dr. Olivier Zava, Dr. Anne Gouget, Romain Dubresson,
Michael, Raphaël Terrier, Farid Mosbaoui, Myriam Scaffidi, Roberta Lopes-Ventura,
Letitia Ischer, Noa Ferrari. In Heidelberg: Dr. Julia Bubeck, Dr. Markus Langhans, Dr.
Giselbert Hinz, Dr. Peter Pimpl, David Scheuring, Silke Niemes, Christina Larenz,
Barbara Jesenofsky, Steffie Gold, Dr. Corrado Viotti, Dr. Stefan Hillmer, Sarah
Colanesi, Nicole Frey, Dr. Peter Oliviuson, Dr. Susanne Holstein, Eva Besemfelder-
Butz. In Milan: Dr. Maddalena de Virgilio, Dr. Alessandra Barbante, Dr. Anna Giulini,
Dr. Emanuela Pedrazzini, Marie Maîtrejean, Andrea Pompa, Giusseppe, Dr. Angelo
Viotti, Dr. Aldo Cerotti, Dr. Floriana Gavazzi, Dr. Diego Breviario, Dr. Silvia Giani,
Ida Brambilla, Flavia Branfi, Antonia Allevi for their kind advice and friendliness.
My thanks to the gardener in Heidelberg, Mr. Michael Schilbach, who took care of my
tobacco plants. I may also thank the tobacco plants and the different bacteria I used
during this work as without them I could not achieve any results.

I wish to thank my family and friends. The people I have met while doing my PhD
rotation and have become my closest and dearest friends and counsellors, and to all of
you I give my love and thanks. In Neuchâtel: Elena Morales, François Zurcher,
Doramys and Gregor Hodel, Anita Ivanova, Marc Creus, Ana Slaughter, Marco
d’Alessandro, Cécile Fatteberg. In Heidelberg: Julia Bubeck, Nuno Santos, Vitor Vieira,
Arunkumar Kota, Alessandro de Mauro, Magdalena Dyniszuk, Roberto Goya, Tiago de
Oliveira, Juraj Paule, Valeria Pettorino, Iain Brown, Clarissa Falcao, Svitlana
Zhukovska, Chris Byrnes, Juraj Paule, Noemí León, Nicole Schädlich, Katja Batschulat,
Pia Oepen. In Milan: Cristina Cristilli, Marie Maîtrejean, Sara, Alessandra Barbante,
Maddalena de Virgilio, Filippo, Antonella, Floriana, Monica, Alessio.
In Catalonia and around the world: Alicia Montes, Laura López, Anna Rogero, Xavi
Ramírez, Jordi Picas, Maria Laroque, Irene Puga, Corinne Pubill, Marina Castellarnau,
María Martín, Elías Pezzat, Juanjo Moreno, Lilian Li, Armin Djamei, Omar and Norma
Sued, Harold Oliva and Carla Ferrada, Núria Climent, Chema Martínez, Massimiliano
Molinari and Jessica Beckett, Mariarosa Mettifogo and Diego Buja.

I would like to give a special thank to my beloved parents, Miquel-Àngel and Neus,
who have always believed in me and helped me to reach my goals. Their unconditional
support forged my desire to achieve all that I wish in life. I owe them everything and
wish I could show them just how much I love and appreciate them. My sister Laia, who
has always encouraged me in my journeys “outside home” and for her inconditional
love and trust. My boyfriend Jordi, whose love and encouragement allowed me to finish
this journey, and already has my heart and love. Finally, I would also like to dedicate
this work to my beloved grandfather Antonio and to the departed soul of my beloved
grandmother Magdalena. I hope that this work makes you proud. Index
INTRODUCTION........................................................................................................................... 1
I Molecular farming - plants as expression system............................................................ 2
II Factors affecting recombinant protein expression .......................................................... 3
II.1 Selection of the promoter..............................................................................................................3
II.2 Codon usage .................................................................................................................................4
II.3 Glycosylation................................................................................................................................5
II.4 Subcellular protein targeting........................................................................................................6
III Choice of the host plant................................................................................................... 9
III.1 Leafy crops...............9
III.2 Cereals and legumes...................................................................................................................10
III.3 Fruits and vegetables .................................................................................................................12
III.4 Fibre and oil crops.........15
IV Plant transformation ...................................................................................................... 15
IV.1 Stable transformation.......15
IV.1.1 Nuclear transformation.........................................................................................................15
IV.1.2 Chloroplast transformation ...................................................................................................16
IV.2 Transient transformation............................................................................................................17
IV.2.1 Agrobacterium-mediated transient expression (agroinfiltration)..........................................17
IV.2.2 Protoplasts ............................................................................................................................17
IV.2.3 Virus-mediated .....................................................................................................................18
V Strategies for recombinant protein accumulation.......................................................... 18
V.1 Membrane proteins.....................................................................................................................19
V.1.1 Types of membrane proteins ...........................................................